LOCUS       sequence-430811-        6671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The DuET construct encodes the named human GPCR engineered with a 
            cleaved hemagglutinin signal peptide, a 5' FLAG and a 3' 1D4 epitope
            tag optimized for expression in mammalian cells and proteomics 
            studies..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Dahl L, Kotliar IB, Bendes A, Dodig-Crnkovic T, Fromm S, Elofsson A,
            Uhlen M, Sakmar TP, Schwenk JM
  TITLE     Multiplexed selectivity screening of anti-GPCR antibodies.
  JOURNAL   Sci Adv. 2023 May 3;9(18):eadf9297. doi: 10.1126/sciadv.adf9297. 
            Epub 2023 May 3.
  PUBMED    37134173
REFERENCE   2  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2023-05-3"; volume: "9(18):eadf9297. doi"; pages: " 
            10.1126/sciadv.adf9297. Epub 2023 May 3"
COMMENT     Sequence Label: GPR120-DuET
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(653..757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(832..851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1023..1402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1403..1606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1557..1577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1651..1670
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1651..1669
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3050..3067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3056..3280
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3326..3754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3413..3432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3623..3644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3763..3783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3768..4097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3948..4083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4010..4029
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4164..4958
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4218..4237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4828..4847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5132..5253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5169..5188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5223..5242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5302..5318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5302..5318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5315..5337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5326..5342
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5350..5356,5357..5374,5375..5380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5395..5416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5533..5550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5784..5803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6463..6671,1..583,584..652))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
       61 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      781 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
      841 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
      901 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca tttgcatgaa
      961 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
     1021 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     1081 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     1141 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     1201 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     1261 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     1321 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     1381 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     1441 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1501 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1561 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
     1621 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
     1681 tggctagcgt ttaaacttaa gcttagcgcc accatgaaga cgatcatcgc cctgagctac
     1741 atcttctgcc tggtattcgc cgactacaag gacgatgatg acgccagcat cgatatgtct
     1801 cctgagtgcg cgagggctgc tggagacgca cccctgaggt cccttgagca agccaaccgc
     1861 actagattcc cctttttctc tgacgtcaag ggtgaccaca ggctggtgct tgcagccgta
     1921 gagactacgg tcctcgttct gattttcgcc gttagtctct tggggaatgt gtgtgctctg
     1981 gtcctggttg caaggcggag gcgcagaggc gccaccgcat gcctcgtgtt gaatctgttt
     2041 tgtgccgatc tcctcttcat tagcgccatt cctctcgtcc tggccgtgcg ctggaccgaa
     2101 gcttggctgc tgggccccgt ggcttgtcac ctccttttct atgttatgac cctgtctggt
     2161 tcagtgacaa tcctgaccct tgcagccgtg tccctggaga ggatggtctg catcgttcat
     2221 cttcaaagag gtgtgcgggg ccctggcagg agggcccggg ccgttctctt ggctcttatc
     2281 tggggatact ctgcagtcgc cgccctcccc ctttgtgtct tttttcgcgt cgtgccacag
     2341 cggctccctg gcgcagacca agaaattagc atttgcaccc tcatttggcc tacgatcccc
     2401 ggtgagattt catgggatgt gtccttcgtg accctgaatt ttttggtacc tggccttgtg
     2461 attgtgatct cttacagcaa aatcctccag accagcgagc acctcctgga tgccagggct
     2521 gtggtgactc actccgaaat taccaaggct agtcgcaaga ggctgaccgt gtccctcgct
     2581 tattcagaaa gtcaccagat tcgcgtgtca cagcaggact ttagactttt taggacgctg
     2641 tttttgctga tggtgagttt ctttattatg tggtccccca ttattattac aatcctgctg
     2701 attcttatcc agaattttaa acaggatctt gtgatctggc ccagcctgtt tttctgggtg
     2761 gtggcattca ctttcgctaa ctcagccctc aatcccatcc tctataatat gacgctttgc
     2821 agaaacgagt ggaagaagat cttctgctgc ttctggtttc cagagaaagg cgctatcctg
     2881 actgatacaa gtgtgaaacg aaatgacctg tctattataa gcgggatcga taccggtgga
     2941 cgcaccggtg aattcgatga ggccagcacc accgtgtcta agaccgagac ttctcaggtg
     3001 gcccctgcct aactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     3061 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3121 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3181 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3241 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3301 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3361 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3421 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3481 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3541 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3601 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3661 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3721 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4021 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4081 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4141 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4201 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4261 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4321 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4381 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4441 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4501 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4561 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4621 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4681 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4741 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4801 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4861 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4921 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4981 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5041 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5101 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5161 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5221 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5281 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaagaacggt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga a
//
